Fig. 2

Overexpression of PVRL4 reduces infections of different types of enveloped viruses in vitro. (A) HEK293T cells were transfected with PVRL4-HA or HA-expressing plasmids. At 24 h post-transfection, the expression of PVRL4-HA was analyzed by western blotting assay. (B-F) PVRL4 or control vector-transfected HEK293T cells were infected with VSV (MOI = 0.01). The GFP were visualized by fluorescence microscopy (B). The GFP-positive cells were analyzed by flow cytometry (C), and the percentages of GFP-positive cells were normalized to the control sample (D). The viral M RNA level and vitral titers in the supernatant was measured by qRT-PCR (E) and plaque assay (F), respectively. Scale bar, 100 μm. The “Lg” means “Log₁₀ change”. (G-K) HEK293T cells were transfected with PVRL4 or vector plasmid for 24 h. Then the cells were infected with HSV-1 (MOI = 0.01) and the GFP was visualized by fluorescence microscopy (G). The GFP-positive cells were analyzed by flow cytometry (H), and values calculated and normalized in (I). The luciferase activity of cells was detected for the indicated times and normalized to the control (J). And after 48 h infection, the vitral titers in supernatants were determined by plaque assay (K). Scale bar, 100 μm. (L-O) HEK293T cells were transfected with PVRL4 or vector plasmid, then these cells were infected with IAV (MOI = 0.01) at 24 h post transfection. After 48 h infection, the viral NP vRNA, mRNA and cRNA level were measured by qRT-PCR (L-N), and viral NP protein expression were measured by western blotting assay (O). Mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001